CN110420330A - A kind of PI3K and MTH1 targeting drug composition and its application - Google Patents
A kind of PI3K and MTH1 targeting drug composition and its application Download PDFInfo
- Publication number
- CN110420330A CN110420330A CN201910653369.5A CN201910653369A CN110420330A CN 110420330 A CN110420330 A CN 110420330A CN 201910653369 A CN201910653369 A CN 201910653369A CN 110420330 A CN110420330 A CN 110420330A
- Authority
- CN
- China
- Prior art keywords
- cell
- bkm120
- pi3k
- medicine
- mth1
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
A kind of PI3K and MTH1 targeting drug composition and its application.Small molecule targeted medicament composition and pharmaceutical preparation provided by the invention, the first preparation formed including the carrier of PI3K inhibitor or its pharmaceutical acceptable, and the second preparation of the carrier formation of MTH1 inhibitor or its pharmaceutical acceptable, compatibility is reasonable, the two drug combination can improve the drug susceptibility of PI3K inhibitor, MTH1 micromolecular inhibitor can be played again in the effect of anti-tumor aspect, increase the therapeutic effect of PI3K inhibitor.The experiment proved that, inhibit significant to activity of tumor cells when both PI3K inhibitor and MTH1 inhibitor drug combination are compared with independent medication to reinforce, and reduce independent medication dosage, drug toxicity can be reduced, it is expected to become the therapeutic scheme new for the undesirable tumour of PI3K inhibitor for treating effect, has broad application prospects.
Description
Technical field
The invention belongs to pharmaceutical technology field more particularly to two kinds of PI3K micromolecular inhibitor BKM120 and GDC0941 and
Two kinds of MTH1 micromolecular inhibitor TH588 and TH287 combination medicines are inhibiting the application in growth of tumour cell, especially exist
Preparation inhibits the application in glioma and lung cancer cell growth drug.
Background technique
With the fast development of human social economy, the disease incidence of malignant tumour is also constantly being increased, and seriously threatens people
The life and health of class causes heavy burden to society and family, therefore explores the pathogenesis of malignant tumour, finds effective
Therapy target and therapeutic agent are of great significance.It is shown according to cancer statistical report in 2018, the new hair of national malignant tumor estimation
Sick rate number 380.4 ten thousand (male 211.4 ten thousand, women 169.0 ten thousand), it is average to be diagnosed as cancer more than 10,000 people daily, often
Minute has 7 people to be diagnosed as cancer.According to morbidity number of cases ranking, lung cancer occupies national the first, annual morbidity about 78.1 ten thousand of falling ill,
Thereafter gastric cancer, colorectal cancer, liver cancer and breast cancer are followed successively by.The World Health Organization has issued new common Lung Cancer Types for 2004
Disease incidence is followed successively by lung adenocarcinoma, squamous carcinoma, Small Cell Lung Cancer and maxicell lung cancer from high to low, shared by these variety classes lung cancer
According to disease incidence be followed successively by 31.5%, 29.4%, 17.8%, 9.2%.Lung cancer is as a kind of the pernicious swollen of high incidence high mortality
Tumor, early diagnosis and therapy are of great significance to extension patient survival, raising life in patients.
Glioma is a kind of most common Primary intracranial tumor, according to brain tumor registration center, the U.S. (Central
Brain tumor registry of the United States, CBTRUS) data, in pernicious central nerve neuroma
47% is Gliblastoma (GBM), is one of the tumour that prognosis is worst in general tumour, has invasion strong, progress is fast
The characteristics of.It is at present maximum operation excision combined radiotherapy and chemotherapy again for the standard care of Gliblastoma.
And blood-brain barrier can be penetrated and to the effective chemotherapeutics Temozolomide of Gliblastoma by being uniquely clinically proven
(Temozolomide, TMZ) is also only capable of patient's median survival interval extending to 14.1 months from 12 months.Observation TCGA (
The Cancer Genome Atlas) the Gliblastoma Genome Atlas drawn, it is found that 88% brain colloid is female thin
Gene mutation, including various RTK(Receptor Tyrosine Kinase occur for RTK/RAS/PI3K signal path in born of the same parents' tumor)
Amplification and mutation, by the PI3K(encoding gene of PIP2 phosphoric acid chemical conversion PIP3 be PI3KCA) activity be mutated or pass through dephosphorization
Acidification reverses the inactivating mutation of the Anti-oncogene PTEN of this reaction and the devitalized of negative regulation RAS signal access NF1 to dash forward
Become the abnormal up-regulation that all will lead to PI3K-AKT-mTOR access to promote growth of tumour cell, proliferation.
BKM120 is I class PI3K micromolecular inhibitor, and acting on p110 α/β/δ/γ IC50 is respectively 52nM/
166nM/116nM/262nM can be reduced downstream effect factors A KT's in the tumour of PI3K-AKT-mTOR signal path activation
Phosphorylation, so that tumour cell be inhibited to rise in value.BKM120 is as a kind of PI3K inhibitor in PI3K signal path activated mutant
There is a large amount of clinical research in the tumours such as breast cancer, non-small cell lung cancer, oophoroma.Therefore glioma is as a kind of PI3K high
The tumour of frequency variability, BKM120, which is used as potential research drug, has important clinical meaning.In the present invention, we use different
Concentration BKM120 handles different brain glioblastoma cell systems, and discovery BKM120 can suppress AKT signal path in 1 μM of concentration
Activation, and the BKM120 of 1 μM of concentration can inhibit most of brain glioblastoma cell system to be proliferated in long term growth experiments.But
In the U251 cell line of PI3K signal path activated mutant, 1 μM of concentration BKM120 can significantly lower the expression of p-AKT, but thin
Born of the same parents' proliferation is not inhibited but, and for such brain glioblastoma cell, we utilize 601 kinds of 178 target spots of targeting
Every kind of drug in library and 1 μM of BKM120 are combined, find another and BKM120 is thin in U251 by small-molecule drug library
The MTH1 small molecule targeted drug TH588 for having synthetic lethal to act in born of the same parents system, the combination of two medicines can significantly inhibit U251 cell Proliferation.
Other than brain glioblastoma cell, we have also verified BKM120 and TH588 in other two plants of lung cancer cell lines PC9 and H460
Coordinate repression.GDC-0941 is another effective PI3K α/δ inhibitor, and IC50 is 3 nM in Cell free assay,
There is the selectivity of appropriateness to p110 β (11 times) and (25 times) of p110 γ, we exist GDC-0941 with the TH588 filtered out
Drug combination in U251 cell, discovery also have significant coordinate repression.
TH588 is a kind of effective as selective MTH1 inhibitor, and IC50 5nM, in vitro in research, TH588 can be swollen
Induced DNA damage in oncocyte, and the dead response for causing ATM-p53 to mediate and DNA are repaired;TH588 is found in In vivo study
Tumour growth can be significantly inhibited in SW480, MCF7 and the xenograft tumor Mice Body of carrying patient BRAF V600E mutation.
MTH1(MutT homolog 1) it is used as a kind of pyrophosphatase, the oxidation nucleotide in nucleotide pond can be efficiently cleared up, thus
Protect the cancer cell of fast breeding from high-level ROS bring fatal damage, but normal cell when lacking MTH1 enzyme according to
It is old to be survived by other subpath DNA plerosis oxidative damages.The MTH1 necessity in normal cell and cancer cell
Significant difference to target this therapy of MTH1 treating cancer, and we are by big data analysis, find brain
MTH1 protein expression level in glioma cell is significantly higher than normal cell.
We are by 601 kinds every kind of small molecule targeted drug in drug libraries with 10 μM of concentration and 1 μM of concentration in the present invention
BKM120 combination, filters out a kind of MTH1 inhibitor TH588, finds 1 μM of concentration BKM120 and 10 μM of concentration TH588 in U251
There is significant coordinate repression, grope using concentration gradient, finds 1 μM of concentration BKM120 and 2 μM of concentration TH588 more
Kind brain glioblastoma cell system includes that stronger coordinate repression is shown in U251, and medicine more mono- than BKM120 and TH588 is to U251
Cell growth inhibition is stronger, and drug concentration is lower, solves U251 cell to the resistance problems of the mono- medicine of BKM120, reduces
Use the cytotoxicity of high concentration list medicine.TH287 is another effective as selective MTH1 inhibitor, IC50 0.8nM, we
BKM120 and TH287 is used in combination in U251 cell line, also shows significant coordinate repression.In collaboration inhibition machine
We have found that PI3K inhibitor and MTH1 inhibitor can inhibit swollen by inducing apoptosis of tumour cell and DNA damage in the research of system
Tumor cell growth.
Summary of the invention
The technical issues of solution: the present invention provides a kind of PI3K and MTH1 targeting drug composition and its applications.The medicine
Compositions are PI3K inhibitor and MTH1 inhibitor.Scheme of the present invention specific can inhibit glioma and lung cancer thin
Intracellular growth can improve PI3K inhibitor and inhibit the curative effect in growth of tumour cell.
Technical solution: a kind of PI3K and MTH1 targeting drug composition contains PI3K inhibitor and MTH1 inhibitor.
Preferably, above-mentioned PI3K inhibitor is BKM120 or GDC-0941.
Preferably, above-mentioned BKM120 concentration is 0 to 4 μM, and GDC0941 concentration is 0 to 100 μM;
Preferably, above-mentioned MTH1 inhibitor is TH588 or TH287.
Preferably, above-mentioned TH588 concentration is 0 to 10 μM, and TH287 concentration is 0 to 4 μM.
Above-mentioned PI3K and MTH1 targeting drug composition inhibits the application in growth of tumour cell drug in preparation.
Above-mentioned cancer is glioma or lung cancer.
The utility model has the advantages that the present invention provides a kind of PI3K and MTH1 micromolecular inhibitor composition and pharmaceutical preparation, it is described
PI3K micromolecular inhibitor is BKM120 and GDC0941;MTH1 micromolecular inhibitor is TH588 and TH287;The pharmaceutical preparation
GDC0941 comprising 0 to 4 μM of concentration of BKM120 or 0 to 100 μM of concentration;0 to 10 μM of concentration of TH588 or 0 to 4 μM of concentration
TH287;The two combination can improve the drug susceptibility of PI3K inhibitor and play MTH1 inhibitor in anti-tumor aspect
Effect, collaboration enhance the therapeutic effect of PI3K inhibitor.The experiment proved that BKM120 and TH588 join in U251 cell
With and BKM120 either capable of being replaced with GDC- by inducing cell apoptosis and promoting DNA damage inhibition cell proliferation
TH588 is still replaced with TH287 by 0941, and coordinate repression of two medicines in U251 cell still shows significantly.Exist simultaneously
BKM120 and TH588 coordinate repression still remains in lung carcinoma cell PC9 and H460.Illustrate that MTH1 inhibits in tumour cell
Agent can enhance the therapeutic effect of PI3K inhibitor.It is anti-that micromolecular inhibitor composition and pharmaceutical preparation of the present invention can apply to preparation
In tumour medicine, have broad application prospects.
Detailed description of the invention
Fig. 1 is influence diagram of the various concentration BKM120 to U251, U87, T98G cell-signaling pathways;Various concentration BKM120
Handle GBM cell line for 24 hours after, intracellular phospho-AKT(Ser 473, Thr 308), phospho-P70S6K(Thr
389), phospho-S6(Ser235/236) it is horizontal in concentration gradient decline, illustrate that AKT/mTOR signal path is suppressed.Phase
With under concentration, sensitive cell line U87, T98G inhibit obvious compared with medicine-resistant cell line signal path.
Fig. 2 is influence diagram of the various concentration BKM120 to a variety of brain glioblastoma cell systems long term growth;Except medicine-resistant cell line
Outside U251 and SNB19, the BKM120 of 1 μM of concentration produces significant inhibition to the long term growth of most of brain glioblastoma cell and makees
With.
Fig. 3 is the synergy figure of the BKM120 and TH588 of various concentration combination in a variety of brain glioblastoma cells;
TH588 and BKM120 various concentration combination under, in addition to T98G cell line, SNB19, LN18, LN229, A172, U251,
There is coordinate repression in U118MG cell line.
Fig. 4 is thin in a variety of gliomas for the mono- medicine of BKM120 and TH588 of various concentration and the combination of two medicine various concentrations
To the influence diagram of cell long-period growth in born of the same parents system;The BKM120 and TH588 two in SNB19, LN229, U251, LN18 cell line
Medicine is combined the inhibiting effect that two drug list medicines are significantly stronger than to the inhibiting effect that cell long-period is grown.
Fig. 5 is the influence diagram under the mono- medicine of BKM120 and TH588 and two medicine synergy to Apoptosis;U251 cell warp
After 1 μM of BKM120,2 μM of TH588 handle 72 hours alone or in combination, the more single medicine processing group of two medicine Combined Treatments, cell withers
Die horizontal increase.
Fig. 6 is the shadow under the mono- medicine of comet detection BKM120 and TH588 and two medicine synergy to DNA Damage
Ring figure;U251 cell is after 1 μM of BKM120,2 μM of TH588 handle 24 hours alone or in combination, when two medicine synergy
The ratio that U251 cell trailing length accounts for cell length reaches 9%, and DNA damage is significantly reinforced compared with the effect of single medicine group.
Fig. 7 is under the mono- medicine of flow cytomery BKM120 and TH588 and two medicine synergy to DNA Damage
Influence diagram;U251 cell is after 1 μM of BKM120,2 μM of TH588 handle 24 hours alone or in combination, when two medicine synergy,
The U251 cell number that DNA damage occurs also accounts for sum to reach 52%, U251 DNA Damage degree more stronger than single medicine.
Fig. 8 is two medicine synergy figures of the BKM120 and TH588 in U251 cell line;BKM120 is difference dense with TH588
Degree combination is lower to handle U251 cell 72h, calculates two medicine index of cooperations with Compusyn software, horizontal line indicates CI=1, CI < 1 in figure
When two medicines have synergistic effect;BKM120 and TH588 has coordinate repression in U251 cell line.
Fig. 9 is two medicine synergy figures of the GDC-0941 and TH588 in U251 cell line;Pan-PI3K inhibitor
The combination of GDC0941 and TH588 various concentration is lower to handle U251 cell 72h, calculates two medicine index of cooperations with Compusyn software,
Two medicines have synergistic effect when horizontal line expression CI=1, CI < 1 in figure;GDC0941 and TH588 has collaboration in U251 cell line
Inhibiting effect.
Figure 10 is two medicine synergy figures of the BKM120 and TH287 in U251 cell line;BKM120 and MTH1 inhibitor
The combination of TH287 various concentration is lower to handle U251 cell 72h, calculates two medicine index of cooperations, horizontal line table in figure with Compusyn software
Show CI=1, two medicines have synergistic effect when CI < 1;BKM120 and TH287 has coordinate repression in U251 cell line.
Figure 11 is two medicine synergy figures of the BKM120 and TH588 in H460 cell line;BKM120 is difference dense with TH588
Degree combination is lower to handle H460 cell 72h, calculates two medicine index of cooperations with Compusyn software, horizontal line indicates CI=1, CI < 1 in figure
When two medicines have synergistic effect;BKM120 and TH588 has coordinate repression in lung cancer H460 cell line.
Figure 12 is two medicine synergy figures of the BKM120 and TH588 in PC9 cell line.BKM120 is difference dense with TH588
Degree combination is lower to handle H460 cell 72h, calculates two medicine index of cooperations with Compusyn software, horizontal line indicates CI=1, CI < 1 in figure
When two medicines have synergistic effect;BKM120 and TH588 has coordinate repression in lung cancer PC9 cell line.
Specific embodiment
The following examples can make those skilled in the art that the present invention be more fully understood, but not limit this in any way
Invention.
It is an object of the invention to find the drug for being directed to the drug resistant tumour cell of PI3K inhibitor, improve PI3K inhibitor
Curative effect in treatment tumour, improves the life cycle of tumor patient.Research shows that can achieve when different Drug combination
The effect that multiple target point inhibits more, in the treatment of malignant tumour, drug combination has become a kind of trend.Different is medication combined
Using certain synergistic effect may be generated, to reduce the dosage of independent medication, the accumulation of drug toxicity in vivo is reduced,
The superposition for avoiding adverse reaction increases the curative effect of drug.
This research starts to select U251 cell line to be higher because of PI3K activated form mutation incidence in glioma, and
BKM120 is as a kind of PI3K inhibitor in the breast cancer of PI3K signal path activated mutant, non-small cell lung cancer, oophoroma etc.
There is a large amount of clinical research in tumour, future may be applied to the treatment of PI3K activated form mutated tumor.We expect BKM120
The brain glioblastoma cell of all PI3K activated form mutation can be inhibited to grow, but experiments have shown that the brain being mutated in PI3K activated form
In glioma cell of U251,1 μM of concentration BKM120 can effectively inhibiting rate AKT signal path be activated, but cannot inhibit cell
Proliferation, therefore guess that also unknown signal path has U251 cell Proliferation with some other than relying on the activation of PI3K signal path
It closes.Then we target the drug libraries of 178 target spots, by every kind of medicine in library using 601 kinds of small molecule targeted drugs are contained
Object is combined with 10 μM of concentration and 1 μM of concentration BKM120, is found one kind and is made with BKM120 with synthetic lethal in U251 cell line
MTH1 inhibitor TH588.Grope by concentration gradient research, it is found that 1 μM of concentration BKM120 and 2 μM of concentration TH588 are combined,
It can inhibit most of brain glioblastoma cell proliferation, the U251 cell line of growth can not be inhibited including the mono- medicine of BKM120.And
Find that the combination of two medicine of BKM120 and TH588 is to promote DNA damage to inhibit U251 by inducing cell apoptosis by Mechanism Study
Cell Proliferation.BKM120 is either replaced with GDC-0941, TH588 is still replaced with TH287, two medicines are in U251 cell
In coordinate repression still show significantly.BKM120 and TH588 collaboration inhibits to make in lung carcinoma cell PC9 and H460 simultaneously
With still remaining.Illustrate that MTH1 inhibitor can enhance the therapeutic effect of PI3K inhibitor in tumour cell.Small molecule of the present invention
Inhibitor combination and pharmaceutical preparation can apply to prepare in anti-tumor drug, have broad application prospects.
The present invention provides a kind of PI3K inhibitor and MTH1 inhibitor combination and pharmaceutical preparation to prepare antineoplastic
Application in object.
BKM120 is a kind of PI3K micromolecular inhibitor, and acting on p110 α/β/δ/γ IC50 is respectively 52nM/
166nM/116nM/262nM can be reduced downstream effect factors A KT's in the tumour of PI3K-AKT-mTOR signal path activation
Phosphorylation, so that tumour cell be inhibited to rise in value.BKM120 is as a kind of PI3K inhibitor in PI3K signal path activated mutant
There is a large amount of clinical research in the tumours such as breast cancer, non-small cell lung cancer, oophoroma.Therefore glioma is as a kind of PI3K high
The tumour of frequency variability, BKM120, which is used as potential research drug, has important clinical meaning.GDC-0941 is that another kind has
PI3K α/δ inhibitor of effect, IC50 is 3 nM in Cell free assay, is had to p110 β (11 times) and p110 γ (25 times)
The selectivity of appropriateness, by GDC-0941 and the TH588 filtered out, the drug combination in U251 cell, discovery also have significantly for we
Coordinate repression.
TH588 is a kind of effective as selective MTH1 inhibitor, and IC50 5nM, in vitro in research, TH588 can be swollen
Induced DNA damage in oncocyte, and the dead response for causing ATM-p53 to mediate and DNA are repaired;TH588 is found in In vivo study
Tumour growth can be significantly inhibited in SW480, MCF7 and the xenograft tumor Mice Body of carrying patient BRAFV600E mutation.
TH287 is another effective as selective MTH1 inhibitor, and IC50 0.8nM, we are used in combination in U251 cell line
BKM120 and TH287 also shows significant coordinate repression.
The present invention source of BKM120, GDC-0941 and TH588, TH287 are not done it is specifically limited, using art technology
Known to personnel, as prepared by commercial source, or use this field routine techniques.
Further, the first preparation and MTH1 that the carrier of the PI3K inhibitor and pharmaceutical acceptable is formed inhibit
The second preparation that the carrier of agent and pharmaceutical acceptable is formed.
Further, the BKM120 concentration is 0 to 4 μM, and GDC0941 concentration is 0 to 100 μM;TH588 concentration is 0
To 10 μM, TH287 concentration is 0 to 4 μM.
The present invention also provides the auxiliary materials of the pharmaceutical composition and pharmaceutical acceptable.
Preferably, the dosage form of the pharmaceutical preparation is injection, tablet, capsule, granule, suspension, emulsion, solution
Agent, glue, freeze drying powder injection, mucilage, aerosol, micro-capsule, microballoon, liposome, micella, sustained release preparation or controlled release preparation.
The present invention also provides said medicines and combination medicine to prepare the application in anticancer drug.
Preferably, the cancer is mainly glioma and lung cancer.
Clear, complete description is carried out to technical solution of the present invention below in conjunction with description of the invention attached drawing, it is clear that
Described embodiment is a part of the embodiment of the present invention, instead of all the embodiments.Those skilled in the art should manage
Solution, modifies to specific embodiments of the present invention or is replaced on an equal basis to some technical characteristics, without departing from the present invention
The spirit of technical solution should all cover in the scope of protection of the invention.
Embodiment 1
It is tied up under the mono- medicine processing of various concentration BKM120 using Western blot detection U251, T98G, U87 brain glioblastoma cell
The variation of signal path, specific embodiment are as follows:
1, U251, T98G, U87 cell line are mentioned into the previous day by 6 X 105/ ware is layered in 6cm ware, and every kind of cell spreads 7 wares.
2,0.1 μM, 1 μM, 2 μM, 4 μM, 8 μM, 10 μM of concentration of list medicine BKM120 is prepared.
3, the drug in 5mL step 2 is added in every ware for 24 hours after cell is completed, and control group adds the DMSO containing 2 ‰ concentration, locates
Reason is for 24 hours.
4, drug-treated is for 24 hours afterwards washed cell twice with PBS, is drained, and 200 μ L cell pyrolysis liquids are added in every ware, uses sleaker
Cracked cell is collected into 1.5mL EP pipe, then in 4 DEG C of cracking half an hour, 13000rpm centrifugation 15min takes supernatant, protects
It is stored in -80 DEG C.
5, each sample protein content is measured with BCA method.
6, the polyacrylamide gel for preparing 10% concentration is used for Protein Separation, and protein sample every hole loading 20ug, 100V are permanent
Piezoelectricity swimming, until bromophenol blue disappears, 250mA is wet to turn 2h, and 3%BSA closes 3 hours, and primary antibody 4 DEG C of incubations 6h, 1 X TBST wash film 3
Secondary, each 10min, secondary antibody is incubated at room temperature 1h, and 1 X TBST is washed film 5 times, each 6min, the colour developing exposure of ECL developing solution.
As shown in Figure 1, when BKM120 concentration is 1 μM, under p-AKT (Ser473) expression is significant in U251, U87, T98G
Drop, shows that PI3K activity is effectively suppressed;And 1 μM of concentration BKM120 can effectively inhibit the expression of p-s6, grow to cell
Also important inhibiting effect should be produced.
Embodiment 2
A variety of brain glioblastoma cells are detected using colony formation and tie up to long term growth situation under the mono- medicine of BKM120 acts on, specifically
Embodiment is as follows:
1, by 8 kinds of brain glioblastoma cell system U251, LN18, SNB19, U118, LN229, U87, A172, T98G with 800, every hole
The amount kind of cell is in 12 orifice plates.
2, the BKM120 of 500nM, 750nM and 1 μM of concentration are prepared, and one group of 2 ‰ concentration DMSO of blank control is set.
3, adherent to the cell in step 1 after 48h, every hole adds the drug prepared in 2mL step 2, and dressing in 3 days is primary, even
After continuous culture 14 days, withdrawal 3 days.
4, withdrawal removes culture medium after 3 days, and PBS is washed 2 times, and deionization is washed 1 time, the fixed 15min of 4% paraformaldehyde, go from
Son washing 1 time, 1 × Ji nurse Sa dye liquor are protected from light dyeing 30min, and deionization is washed 3 times, preservation of taking pictures.
As shown in Fig. 2, the BKM120 of 1 μM of concentration is to most of brain glioblastoma cell in addition to U251 and SNB19 cell line
Long term growth produces significant inhibiting effect.
Embodiment 3
Lower cell proliferative conditions are combined using the mono- medicine of Alamar blue detection BKM120, TH588 and two medicines, and calculate SI
Value, specific embodiment are as follows:
1, the previous day by U251, SNB19 according to 1000 cell per well kinds in 96 orifice plates;LN229,A72,LN18,U118MG,
For T98G by 2000 cell per well kinds in 96 orifice plates, each concentration of every kind of cell spreads 3 multiple holes, and control group is arranged.
2, single medicine BKM120 100nM, 1 BKM120 μM, 10 μM of TH588,2 μM of TH588 are prepared;Double medicine BKM120
100nM+ TH588 10μM、BKM120 100nM+ TH588 2μM、BKM120 1μM+ TH588 10μM、BKM120 1μM+
TH588 2μM。
3, it is inhaled after cell is adherent in step 1 and abandons old culture medium, then 100 μ L complete mediums and 10 μ L are added in every hole
The mixed liquor of Alamar Blue is existed after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide using Chemiluminescence Apparatus
Absorbance is detected under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data, is recorded as Day0 number
According to.
4, the cell for having surveyed Day0 data is inhaled to the mixed liquor for abandoning the Blue containing Alamar, then every hole is added what step 2 was prepared
Drug, third day Alamar Blue detect cell growth, and method such as step 2 is recorded as Day3 data.
5, the mono- medicine of TH588 is acted on into lower cell survival rate and is denoted as Rc;The mono- medicine of BKM120 acts on lower cell survival rate and is denoted as Cd;
Cc will be denoted as without cell survival rate under drug effect;Cell survival rate in the case of the combination of two medicines is denoted as Rd;ECE = (Rc/Cc)
× (Cd/Cc);OCE = Rd/Cc;SI = ECE - OCE;As SI > 0, the combination of two medicines has synergistic effect;When SI < 0
When, the combination of two medicines has antagonism.According to the data of Day3 days records, every kind of cell SI value is calculated and such as the Fig. 3 that maps.
As shown in figure 3, when BKM120 concentration is 1 μM, two medicines are in SNB19, LN229, A172 when TH588 concentration is 10 μM
And there is coordinate repression in LN18 cell line;When TH588 concentration is 2 μM, and BKM120 concentration is 1 μM, two medicines exist
There is coordinate repression in SNB19, LN229, A172, U251, U118MG cell line;TH588 concentration is 2 μM, and BKM120 is dense
It is 10 μM that degree, which is 100nM and TH588 concentration, and BKM120 concentration is for 100nM all without coordinate repression in T98G.So
1 μM of BKM120 and 2 μM of TH588 is brain glioblastoma cell system optimum synergistic inhibiting effect concentration.
Embodiment 4
Using colony formation detect a variety of brain glioblastoma cells tie up to the mono- medicine of BKM120, the mono- medicine of TH588 and BKM120 with
A variety of brain glioblastoma cell long term growth situations in the case of TH588 combination, specific embodiment are as follows:
1, mention the previous day by U251, SNB19 cell line by 1000 cell per well kinds in 12 orifice plates, A172, U118MG,
LN18, LN229 and T98G are by 2000 cell per well kinds in 12 orifice plates.
2, prepare list medicine BKM120 100nM, 1 μM of single medicine BKM120,2 μM of single medicine TH588,1 μM of two medicine BKM120+
TH588 2μM;Two 2 μM of medicine BKM120 100nM+TH588.
3, after the cell in step 1 is completely adherent, the drug in step 2 is added in adherent cell, is changed every three days
Liquid is primary, and dosing culture 14 days, 3 days poststainings of withdrawal.
4, withdrawal removes culture medium after 3 days, and PBS is washed 2 times, and deionization is washed 1 time, the fixed 15min of 4% paraformaldehyde, go from
Son washing 1 time, 1 × Ji nurse Sa dye liquor are protected from light dyeing 30min, and deionization is washed 3 times, preservation of taking pictures.
As shown in figure 4, BKM120 and the combination of TH588 two medicine grow cell long-period in SNB19, LN229, U251 cell line
Inhibiting effect is significantly stronger than the inhibiting effect of two drug list medicines.
Embodiment 5
Using the U251 Apoptosis situation under Annexin V-FITC detection BKM120 and the mono- medicine of TH588 and double medicine effects.
Specific embodiment is as follows:
1, by U251 cell according to 4 × 104Control group, the mono- medicine group of BKM120, the mono- medicine of TH588 is arranged in 6 orifice plates in a every hole kind
Group, two medicine joint group of BKM120+TH588;3 multiple holes of every group of setting;And flow cytometer is reserved in advance.
2, single 1 μM of medicine BKM120,2 μM of TH588 and 1 μM of BKM120,2 μM of+TH588 are prepared.
3, the culture medium in step 2 is added after cell is adherent, cultivates 3 days.
4, culture medium in 6cm ware is transferred to completely in 15mL centrifuge tube, PBS is washed twice of cell, and is transferred to 15mL
Centrifuge tube.
5,300 μ L pancreatin are added, after shaking up, quickly sops up 200u pancreatin and (guarantees to be covered by pancreatin on cell, avoid
Pancreatin containing EDTA to) terminated with 1mL complete medium, and be transferred to 15mL centrifuge tube, then wash one with 1mL complete medium
Time, go to 1500rpm in centrifuge tube, be centrifuged after 5min to inhale and abandon supernatant, then wash cell with the PBS that 1mL is pre-chilled, 300g, 4 degree from
Heart 5min, is repeated once;It inhales and abandons PBS, 100 μ L 1XBinding buffer are added, cell is resuspended, 2.5 μ L Annexin are added
V-FITC and 5 μ L PI staining Solution, mixes gently;It is protected from light incubation at room temperature 15min, mixes one every 5min
It is secondary;400 μ L 1 × Binding Buffer are added, is placed in after mixing on ice, waits flow cytometer detection (completing in 1 hour) to examination
Data are tested to analyze and count.
As shown in figure 5, only 6.5%, 2 μM of single medicine TH588 are made U251 apoptosis rate when single medicine 1 μM of effect of BKM120
Used time U251 apoptosis rate is 9.5%, but U251 apoptosis rate reaches 18% when two medicine synergy, and two medicine synergy are drawn
Play Apoptosis quantity and be significantly higher than single medicine,p < 0.05 difference is statistically significant.
Embodiment 6
Lower U251 DNA Damage feelings are acted on using the mono- medicine of comet detection BKM120, the mono- medicine of TH588 and two medicine combinations
Condition.Specific embodiment is as follows:
1, by U251 cell line according to 1.5 × 105A every hole kind in 6 orifice plates, the mono- medicine group of setting BKM120, the mono- medicine group of TH588,
Two medicine combination groups, positive controls, negative control group;
2, mono- 1 μM of medicine of BKM120 of preparation, 2 μM of the mono- medicine of TH588, the double medicines of 1 μM of BKM120,2 μM of+TH588,100 μM of H2O2。
3, after the U251 cell in step 1 is completely adherent, changing culture medium into drug in step 2, (positive control is removed
It handles outside) for 24 hours;
4, drug-treated is washed once with 1 × PBS afterwards for 24 hours, then digests counting with pancreatin.The cell counted is washed with PBS again
Once, it is then resuspended with PBS to 1 × 106/mL of concentration.
5, film-making: the normal melting point agarose drop that the PBS of 100 μ L 0.75% is dissolved closes the lid on frosted glass slide
Slide avoids bubble, and 4 DEG C of refrigerator 10min after solidification, carefully remove coverslip, this is first layer glue (lower layer's glue)
6, (temperature is that 40-42 just loses no time to mix to the low melting-point agarose for taking the PBS of 10 μ L cell suspensions and 90 μ L 0.75% to dissolve
It is even) it mixes, it is added dropwise on first layer glue, covered, 4 DEG C of refrigerators stand 30min, and solidification carefully removes coverslip, this is
Second layer glue (upper layer glue)
7, slide cell cracking: is placed in cell pyrolysis liquid (10mM Tris pH=10,2.5M of the good pre-cooling of Fresh
Nacl, 0.1M EDTA2Na, 10% DMSO, 1% Tritox-100) in, it is protected from light, 4 DEG C of cracking are overnight.
8, with enzyme reaction buffer solution (40mM HEPES pH=8.0,0.1M Kcl, 0.5mM EDTA2Na, BSA
It 0.2mg/mL) washes three times;
9, electrophoresis: slide is placed in Horizontal electrophoresis tank, pre-cooling alkaline electrophoresis buffer (10M NaOH (pH=13),
0.5M EDTA2Na) in stand 20min, electrophoresis 25V, electric current 300mA, electrophoresis 30min.
10, it neutralizes and dyes: after electrophoresis, impregnating slide, 2.5ug/mL using neutralizer (0.4M Tris-HCl)
20 μ L of DAPI is protected from light dyeing, 15min;
11, fluorescence microscopy is taken pictures under the microscope (DAPI uses burst of ultraviolel);
12, the ratio that the length that cell trails in every group accounts for the cell whole length is calculated with Casp software, ratio gets over high DNA damage
Hurt bigger.
As shown in fig. 6, U251 cell trailing length accounts for the ratio about 4% of cell length when the mono- medicine of 1 μM of BKM120 acts on;2
When the mono- medicine effect of μM TH588, U251 cell trailing length accounts for the ratio about 5% of cell length;But when two medicine synergy
The ratio that U251 cell trailing length accounts for cell length reaches 9%, and DNA damage is significantly reinforced compared with the effect of single medicine group.p <
0.05, difference is statistically significant.
Embodiment 7
γ H2AX positive cell is detected using Flow Cytometry, U251 DNA Damage situation is detected with this.Specific embodiment party
Case is as follows:
1, by U251 cell according to 1.2 × 104A every hole kind is in 6 orifice plates;The mono- medicine group of BKM120, the mono- medicine group of TH588, two are set
Medicine combination group, negative control group;(reserving flow cytometer in advance);
2, single 1 μM of medicine BKM120, single 2 μM of medicine TH588,1 μM of double medicine BKM120,2 μM of+TH588 are prepared;
3, after cell in step 1 is adherent, the drug in step 2 is added, the hole 3mL/ handles 48h;
4, cell dissociation is centrifuged after drug-treated, removes supernatant, 1mL PBS is resuspended;
5,3mL 90% is taken to analyze alcohol (- 20 DEG C of pre-coolings) in 15mL centrifuge tube, 1mL is added in side vortex (soft, concussion instrument) side
The cell of PBS respin, -20 DEG C of fixations are overnight;
6,1500rpm, 8min remove supernatant, and 1mL FASC respin is simultaneously transferred in 1.5mL EP pipe, and 5000rpm 2min is gone
Clearly, 500 μ L FACS respin, 5000rpm 2min remove supernatant;
7, match antibody processed: 1:50 (5 samples add 5 μ L of antibody, add 250 μ L of FACS, mix, and a sample adds 50 μ L);
8, it is added and matches antibody processed, room temperature, which is protected from light, is incubated for 1h, and 20min is flicked once;
9, add 1mL FACS respin, centrifugation, 5000rpm 2min;
10, supernatant is removed, 500 μ L FACS, 5000rpm 2min are added;
11, plus 300 μ L FACS respins, flow cytometer detect.
As shown in fig. 7, the U251 cell number that DNA damage occurs accounts for the 40% of sum when the mono- medicine of 1 μM of BKM120 acts on;2μM
When the mono- medicine of TH588 acts on, the U251 cell number that DNA damage occurs also accounts for sum about 40%;But when two medicine synergy, hair
The U251 cell number of raw DNA damage also accounts for sum and reaches 52%, and when illustrating two medicine synergy, U251 DNA Damage degree is more
By force.p< 0.05, difference is statistically significant.
Embodiment 8
By multiple concentration BKM120 and multiple concentration TH588 drug combinations, observe whether they have collaboration in U251 cell line
Inhibiting effect.Specific embodiment is as follows:
1, by U251 cell according to 1500, every hole cell kind in 96 orifice plates;
2, concentration gradient, six concentration gradients of every kind of drug are set, and the drug containing that combination of two is made into 36 groups of final concentrations is cultivated completely
Base;
BKM120 ( 0μM、0.5μM、0.75μM、1μM、2μM、4μM)
TH588 ( 0μM、1μM、1.5μM、2μM、4μM、8μM)
3, plating cells discard the old culture medium of drug containing afterwards for 24 hours, replace the fresh drug containing culture of various concentration combination in step 2
Base;
4, the old culture medium of drug containing is discarded after dosing 72h, every hole adds containing 100 μ L complete mediums and 10 μ L Alamar Blue
Mixed liquor, and negative control is set, after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide, using chemiluminescence
Instrument detects absorbance under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data.
5, two medicine index of cooperations are calculated using Compusyn software, then analysis mapping.
Two medicine index of cooperations are calculated using Compusyn software as shown in Figure 8, horizontal line indicates CI=1 in figure, as CI > 1 two
Medicine has antagonism;Two medicines have addition when CI=1;Two medicines have synergistic effect when CI < 1;In figure as the result is shown BKM120 and
TH588 has coordinate repression in U251 cell line.
Embodiment 9
By multiple concentration GDC0941 and multiple concentration TH588 drug combinations, observe whether they have collaboration in U251 cell line
Inhibiting effect.Specific embodiment is as follows:
1, by U251 cell according to 1500, every hole cell kind in 96 orifice plates;
2, concentration gradient, six concentration gradients of every kind of drug are set, and the drug containing that combination of two is made into 36 groups of final concentrations is cultivated completely
Base;
GDC0941 ( 0μM、1μM、2μM、10μM、20μM、100μM)
TH588 ( 0μM、2μM、4μM、5μM、6μM、8μM)
3, plating cells discard the old culture medium of drug containing afterwards for 24 hours, replace the fresh drug containing culture of various concentration combination in step 2
Base;
4, the old culture medium of drug containing is discarded after dosing 72h, every hole adds containing 100 μ L complete mediums and 10 μ L Alamar Blue
Mixed liquor, and negative control is set, after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide, using chemiluminescence
Instrument detects absorbance under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data.
5, two medicine index of cooperations are calculated using Compusyn software, then analysis mapping.
Two medicine index of cooperations are calculated using Compusyn software as shown in Figure 9, horizontal line indicates CI=1 in figure, as CI > 1 two
Medicine has antagonism;Two medicines have addition when CI=1;Two medicines have synergistic effect when CI < 1;In figure as the result is shown GDC0941 and
TH588 has coordinate repression in U251 cell line.
Embodiment 10
By multiple concentration BKM120 and multiple concentration TH287 drug combinations, observe whether they have collaboration in U251 cell line
Inhibiting effect.Specific embodiment is as follows:
1, by U251 cell according to 1500, every hole cell kind in 96 orifice plates;
2, be arranged concentration gradient, 5 concentration gradients of BKM120,6 concentration gradients of TH287, combination of two be made into 30 groups it is dense eventually
The drug containing complete medium of degree;
BKM120 ( 0μM、0.5μM、1μM、2μM、4μM、)
TH287 ( 0μM、0.25μM、0.5μM、1μM、2μM、4μM)
3, plating cells discard the old culture medium of drug containing afterwards for 24 hours, replace the fresh drug containing culture of various concentration combination in step 2
Base;
4, the old culture medium of drug containing is discarded after dosing 72h, every hole adds containing 100 μ L complete mediums and 10 μ L Alamar Blue
Mixed liquor, and negative control is set, after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide, using chemiluminescence
Instrument detects absorbance under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data.
5, two medicine index of cooperations are calculated using Compusyn software, then analysis mapping.
Two medicine index of cooperations are calculated using Compusyn software as shown in Figure 10, horizontal line indicates CI=1 in figure, as CI > 1
Two medicines have antagonism;Two medicines have addition when CI=1;Two medicines have synergistic effect when CI < 1;In figure as the result is shown BKM120 and
TH287 has coordinate repression in U251 cell line.
Embodiment 11
By multiple concentration BKM120 and multiple concentration TH588 drug combinations, observe whether they have collaboration in H460 cell line
Inhibiting effect.Specific embodiment is as follows:
1, by H460 cell according to 2000, every hole cell kind in 96 orifice plates;
2, be arranged concentration gradient, 6 concentration gradients of BKM120,5 concentration gradients of TH588, combination of two be made into 30 groups it is dense eventually
The drug containing complete medium of degree;
BKM120 ( 0μM、0.2μM、0.4μM、0.6μM、0.8μM、1μM)
TH588( 0μM、2μM、3μM、4μM、6μM)
3, plating cells discard the old culture medium of drug containing afterwards for 24 hours, replace the fresh drug containing culture of various concentration combination in step 2
Base.
4, the old culture medium of drug containing is discarded after dosing 72h, every hole adds containing 100 μ L complete mediums and 10 μ L Alamar
The mixed liquor of Blue, and negative control is set, after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide, using chemistry
Light-emitting appearance detects absorbance under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data.
5, two medicine index of cooperations are calculated using Compusyn software, then analysis mapping.
Two medicine index of cooperations are calculated using Compusyn software as shown in figure 11, horizontal line indicates CI=1 in figure, as CI > 1
Two medicines have antagonism;Two medicines have addition when CI=1;Two medicines have synergistic effect when CI < 1;In figure as the result is shown BKM120 and
TH588 has coordinate repression in H460 cell line.
Embodiment 12
By multiple concentration BKM120 and multiple concentration TH588 drug combinations, observe whether they have collaboration to press down in PC9 cell line
Production is used.Specific embodiment is as follows:
1, by PC9 cell according to 2000, every hole cell kind in 96 orifice plates;
2, be arranged concentration gradient, 5 concentration gradients of BKM120,6 concentration gradients of TH588, combination of two be made into 30 groups it is dense eventually
The drug containing complete medium of degree;
BKM120 ( 0μM、0.5μM、0.75μM、1μM、2μM)
TH588( 0μM、2μM、3μM、4μM、8μM、10μM)
3, plating cells discard the old culture medium of drug containing afterwards for 24 hours, replace the fresh drug containing culture of various concentration combination in step 2
Base;
4, the old culture medium of drug containing is discarded after dosing 72h, every hole adds containing 100 μ L complete mediums and 10 μ L Alamar Blue
Mixed liquor, and negative control is set, after being incubated for 4h in the cell incubator of 37 DEG C of 5% carbon dioxide, using chemiluminescence
Instrument detects absorbance under conditions of excitation wavelength 534nm and wavelength of transmitted light 584nm and records every hole data.
5, two medicine index of cooperations are calculated using Compusyn software, then analysis mapping.
Two medicine index of cooperations are calculated using Compusyn software as shown in figure 12, horizontal line indicates CI=1 in figure, as CI > 1
Two medicines have antagonism;Two medicines have addition when CI=1;Two medicines have synergistic effect when CI < 1;In figure as the result is shown BKM120 and
TH588 has coordinate repression in PC9 cell line.
Claims (7)
1. a kind of PI3K and MTH1 targets drug composition, it is characterised in that contain PI3K inhibitor and MTH1 inhibitor.
2. PI3K and MTH1 target drug composition according to claim 1, it is characterised in that the PI3K inhibitor is
BKM120 or GDC-0941.
3. PI3K and MTH1 targets drug composition according to claim 2, it is characterised in that the BKM120 concentration is 0 to 4 μ
M, GDC0941 concentration are 0 to 100 μM.
4. PI3K and MTH1 targets drug composition according to claim 1, it is characterised in that the MTH1 inhibitor is TH588
Or TH287.
5. PI3K and MTH1 targets drug composition according to claim 4, it is characterised in that the TH588 concentration is 0 to 10 μ
M, TH287 concentration are 0 to 4 μM.
6. any PI3K of claim 1-5 and MTH1 targeting drug composition inhibits in growth of tumour cell drug in preparation
Using.
7. application according to claim 6, it is characterised in that the cancer is glioma or lung cancer.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910653369.5A CN110420330B (en) | 2019-07-19 | 2019-07-19 | Pharmaceutical application of PI3K and MTH1 targeted drug composition |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910653369.5A CN110420330B (en) | 2019-07-19 | 2019-07-19 | Pharmaceutical application of PI3K and MTH1 targeted drug composition |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110420330A true CN110420330A (en) | 2019-11-08 |
CN110420330B CN110420330B (en) | 2020-05-29 |
Family
ID=68410127
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910653369.5A Active CN110420330B (en) | 2019-07-19 | 2019-07-19 | Pharmaceutical application of PI3K and MTH1 targeted drug composition |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110420330B (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102740851A (en) * | 2009-10-12 | 2012-10-17 | 霍夫曼-拉罗奇有限公司 | Combinations of a PI3K inhibitor and a MEK inhibitor |
-
2019
- 2019-07-19 CN CN201910653369.5A patent/CN110420330B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102740851A (en) * | 2009-10-12 | 2012-10-17 | 霍夫曼-拉罗奇有限公司 | Combinations of a PI3K inhibitor and a MEK inhibitor |
Non-Patent Citations (2)
Title |
---|
EIKE TATJANA ARISTIZABAL PRADA, ET AL.: ""The MTH1 inhibitor TH588 demonstrates anti-tumoral effects alone and in combination with everolimus, 5-FU and gamma-irradiation in neuroendocrine tumor cells"", 《PLOS ONE》 * |
仇振巍, 岳双柱 等.: ""磷脂酰肌醇3激酶抑制剂BKM120抑制U251神经胶质瘤细胞增殖并促进其凋亡"", 《细胞与分子免疫学杂志》 * |
Also Published As
Publication number | Publication date |
---|---|
CN110420330B (en) | 2020-05-29 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Matarredona et al. | Neural stem cells of the subventricular zone as the origin of human glioblastoma stem cells. Therapeutic implications | |
Calabrese et al. | A perivascular niche for brain tumor stem cells | |
Ahmed et al. | Understanding glioma stem cells: rationale, clinical relevance and therapeutic strategies | |
CN105582014B (en) | For treating the composition and method of leukaemia | |
Hong et al. | Targeting cancer stem cells by using the nanoparticles | |
Cao et al. | Angiocrine factors deployed by tumor vascular niche induce B cell lymphoma invasiveness and chemoresistance | |
Conyers et al. | Liposarcoma: molecular genetics and therapeutics | |
Bayin et al. | Notch signaling regulates metabolic heterogeneity in glioblastoma stem cells | |
Persano et al. | Glioblastoma cancer stem cells: role of the microenvironment and therapeutic targeting | |
Maher et al. | Malignant glioma: genetics and biology of a grave matter | |
Jane et al. | Coadministration of sorafenib with rottlerin potently inhibits cell proliferation and migration in human malignant glioma cells | |
Park et al. | Biology of glioma cancer stem cells | |
Kodama et al. | Expression of platelet‐derived growth factor (PDGF)‐B and PDGF‐receptor β is associated with lymphatic metastasis in human gastric carcinoma | |
Zhang et al. | Targeting role of glioma stem cells for gliobastoma multiforme | |
Ping et al. | Cancer stem cells and their vascular niche: Do they benefit from each other? | |
Eimer et al. | Cyclopamine cooperates with EGFR inhibition to deplete stem-like cancer cells in glioblastoma-derived spheroid cultures | |
Sales et al. | Stem cells and cancer: an overview | |
CN102869361A (en) | N-4 ( - ( ( 3- ( 2 -amino-4 pyrimidinyl) -2 -pyridinyl) oxy) phenyl) -4- (4-methyl-2-thienyl) -1-phthalazinamine for use in the treatment of antimitotic agent resistant cancer | |
Yuan et al. | Modified Jian-pi-yang-zheng decoction inhibits gastric cancer progression via the macrophage immune checkpoint PI3Kγ | |
Flamini et al. | Significance and therapeutic implications of endothelial progenitor cells in angiogenic-mediated tumour metastasis | |
CN104758292B (en) | PD-0332991 is preparing the purposes of prevention drug-resistant tumor drug | |
Shahrabi et al. | Bone marrow blood vessels: Normal and neoplastic niche | |
Wang et al. | CDK4/6 nano-PROTAC enhances mitochondria-dependent photodynamic therapy and anti-tumor immunity | |
CN111035641B (en) | Novel application of GZD824 and pharmaceutically acceptable salts thereof in treatment of diseases | |
Kim et al. | Anti‐angiogenic activity of thienopyridine derivative LCB 03‐0110 by targeting VEGFR‐2 and JAK/STAT 3 Signalling |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |